Cecil Jessica D, O'Brien-Simpson Neil M, Lenzo Jason C, Holden James A, Chen Yu-Yen, Singleton William, Gause Katelyn T, Yan Yan, Caruso Frank, Reynolds Eric C
Oral Health Cooperative Research Centre, Melbourne Dental School, Bio21 Institute, The University of Melbourne, Melbourne, Victoria, Australia.
Department of Chemical and Biomolecular Engineering, The University of Melbourne, Parkville, Victoria, Australia.
PLoS One. 2016 Apr 1;11(4):e0151967. doi: 10.1371/journal.pone.0151967. eCollection 2016.
Highly purified outer membrane vesicles (OMVs) of the periodontal pathogens, Porphyromonas gingivalis, Treponema denticola and Tannerella forsythia were produced using tangential flow ultrafiltration, ultracentrifugation and Optiprep density gradient separation. Cryo-TEM and light scattering showed OMVs to be single lipid-bilayers with modal diameters of 75 to 158 nm. Enumeration of OMVs by nanoparticle flow-cytometry at the same stage of late exponential culture indicated that P. gingivalis was the most prolific OMV producer. P. gingivalis OMVs induced strong TLR2 and TLR4-specific responses and moderate responses in TLR7, TLR8, TLR9, NOD1 and NOD2 expressing-HEK-Blue cells. Responses to T. forsythia OMVs were less than those of P. gingivalis and T. denticola OMVs induced only weak responses. Compositional analyses of OMVs from the three pathogens demonstrated differences in protein, fatty acids, lipopolysaccharide, peptidoglycan fragments and nucleic acids. Periodontal pathogen OMVs induced differential pattern recognition receptor responses that have implications for their role in chronic periodontitis.
使用切向流超滤、超速离心和Optiprep密度梯度分离法制备了牙周病原体牙龈卟啉单胞菌、具核梭杆菌和福赛坦氏菌的高度纯化外膜囊泡（OMV）。冷冻透射电子显微镜和光散射显示，OMV为单脂质双层，模态直径为75至158纳米。在指数生长后期的同一阶段，通过纳米颗粒流式细胞术对OMV进行计数，结果表明牙龈卟啉单胞菌是最丰富的OMV产生菌。牙龈卟啉单胞菌OMV在表达TLR7、TLR8、TLR9、NOD1和NOD2的HEK-Blue细胞中诱导了强烈的TLR2和TLR4特异性反应以及中等程度的反应。对福赛坦氏菌OMV的反应小于对牙龈卟啉单胞菌的反应，而具核梭杆菌OMV仅诱导了微弱反应。对这三种病原体的OMV进行的成分分析表明，它们在蛋白质、脂肪酸、脂多糖、肽聚糖片段和核酸方面存在差异。牙周病原体OMV诱导了不同的模式识别受体反应，这对它们在慢性牙周炎中的作用具有重要意义。
